Division of Applied Life Science (BK21 Program), EB-NCRC and PMBBRC, Gyeongsang National University, Jinju 660-701, Republic of Korea.
Metab Eng. 2011 Nov;13(6):648-55. doi: 10.1016/j.ymben.2011.08.001. Epub 2011 Sep 1.
Sesquiterpenes are important materials in pharmaceuticals and industry. Metabolic engineering has been successfully used to produce these valuable compounds in microbial hosts. However, the microbial potential of sesquiterpene production is limited by the poor heterologous expression of plant sesquiterpene synthases and the deficient FPP precursor supply. In this study, we engineered E. coli to produce α-farnesene using a codon-optimized α-farnesene synthase and an exogenous MVA pathway. Codon optimization of α-farnesene synthase improved both the synthase expression and α-farnesene production. Augmentation of the metabolic flux for FPP synthesis conferred a 1.6- to 48.0-fold increase in α-farnesene production. An additional increase in α-farnesene production was achieved by the protein fusion of FPP synthase and α-farnesene synthase. The engineered E. coli strain was able to produce 380.0 mg/L of α-farnesene, which is an approximately 317-fold increase over the initial production of 1.2 mg/L.
倍半萜是药物和工业的重要原料。代谢工程已成功用于在微生物宿主中生产这些有价值的化合物。然而，由于植物倍半萜合酶的异源表达效果不佳和 FPP 前体供应不足，微生物生产倍半萜的潜力受到限制。在本研究中，我们使用经过密码子优化的α-法呢烯合酶和外源 MVA 途径对大肠杆菌进行了工程改造，以生产α-法呢烯。α-法呢烯合酶的密码子优化提高了合酶的表达和α-法呢烯的产量。增加 FPP 合成的代谢通量可使α-法呢烯的产量增加 1.6 至 48.0 倍。通过 FPP 合酶和α-法呢烯合酶的蛋白融合进一步提高了α-法呢烯的产量。该工程大肠杆菌菌株能够生产 380.0mg/L 的α-法呢烯，比初始产量 1.2mg/L 提高了约 317 倍。
